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43065
Hypoxia Activation IHC Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Hypoxia Activation IHC Antibody Sampler Kit #43065

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Hypoxia Activation IHC Antibody Sampler Kit: Image 1
Confocal immunofluorescent analysis of Hep G2 cells, untreated (left) or treated with cobalt chloride (500 μM, 24 h; right), using HIF-1α (E1V6A) Rabbit mAb (green) and DyLight™ 650 Phalloidin #12956 (red).
Hypoxia Activation IHC Antibody Sampler Kit: Image 2
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus labeled with Glut1 (E4S6I) Rabbit mAb (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #36618 (right, red), GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657 (right, cyan pseudocolor), and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Hypoxia Activation IHC Antibody Sampler Kit: Image 3
Confocal immunofluorescent analysis of Hep G2 cells (left, high-expressing) and A-204 cells (right, low-expressing) using Glut1 (E4S6I) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
Hypoxia Activation IHC Antibody Sampler Kit: Image 4
Western blot analysis of extracts from wild-type, GSK-3α (-/-), and GSK3β (-/-) mouse embryonic fibroblasts (MEFs) using GSK-3β (D5C5Z) XP® Rabbit mAb (upper) and GSK-3α/β (D75D3) XP® Rabbit mAb #5676 (lower). (MEF wild type, GSK-3α (-/-), and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Hypoxia Activation IHC Antibody Sampler Kit: Image 5
Western blot analysis of extracts from various cell types using LDHA (C4B5) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 6
Western blot analysis of extracts from various cell lines and mouse skeletal muscle using PKM2 (D78A4) XP® Rabbit mAb (upper) or GAPDH (14C10) Rabbit mAb #2118.
Hypoxia Activation IHC Antibody Sampler Kit: Image 7
Western blot analysis of extracts from Hep G2 cells untreated (-) or treated with cobalt chloride (CoCl2) (0.1 mM, 4 hr; +), Raji cells untreated (-) or treated with CoCl2 (0.1 mM, 4 hr; +), and U-2 OS cells untreated (-) or treated with DMOG (1 mM, 6 hr; +), using HIF-1α (E1V6A) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Hypoxia Activation IHC Antibody Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using HIF-1α (E1V6A) Rabbit mAb performed on the Leica® BOND™ Rx.
Hypoxia Activation IHC Antibody Sampler Kit: Image 9
Western blot analysis of extracts from various cell types using HIF-1β/ARNT (D28F3) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 10
Western blot analysis of extracts from Hep G2 and MOLT-4 cells using Glut1 (E4S6I) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Hypoxia Activation IHC Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using VHL (E3X9K) Rabbit mAb performed on the Leica BOND Rx.
Hypoxia Activation IHC Antibody Sampler Kit: Image 12
Western blot analysis from wild-type mouse embryo fibroblasts (MEF) (lane 1) or SirT1 knockout mouse embryo fibroblasts (MEF) (lane 2) using SirT1 (1F3) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SirT1 knockout mouse embryo fibroblasts (MEF) confirms specificity of the antibody for SirT1. Wild-type and knockout MEFs were a gift from Wenyi Wei at the Harvard Medical School.
Hypoxia Activation IHC Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded human salivary gland small cell carcinoma using SirT1 (1F3) Mouse mAb (left) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (right).
Hypoxia Activation IHC Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded normal human parathyroid using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded HCT116 cell pellet, wild-type (left, positive) or SirT1 knockout (right, negative), using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 17
Western blot analysis of extracts from various cell lines using p300 (D8Z4E) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, HCT-15 cells are negative for p300 expression.
Hypoxia Activation IHC Antibody Sampler Kit: Image 18
CUT&RUN was performed with 293T cells and p300 (D8Z4E) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across MCL1, a known target gene of p300 (see additional figure containing qPCR data).
Hypoxia Activation IHC Antibody Sampler Kit: Image 19
Immunoprecipitation of GSK-3β from PC-12 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or GSK-3β (D5C5Z) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using GSK-3β (D5C5Z) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using LDHA (C4B5) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using PKM2 (D78A4) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 22
Western blot analysis of extracts from C2C12 cells, untreated (-) or treated with cobalt chloride (CoCl2) (0.1 mM, 4 hr; +), using HIF-1α (E1V6A) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Hypoxia Activation IHC Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor using HIF-1α (E1V6A) Rabbit mAb performed on the Leica® BOND™ Rx.
Hypoxia Activation IHC Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HIF-1β/ARNT (D28F3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Hypoxia Activation IHC Antibody Sampler Kit: Image 25
Immunoprecipitation of Glut1 protein from HuH-6 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Glut1 (E4S6I) Rabbit mAb. Western blot analysis was performed using Glut1 (E4S6I) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as the secondary antibody.
Hypoxia Activation IHC Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using VHL (E3X9K) Rabbit mAb performed on the Leica BOND Rx.
Hypoxia Activation IHC Antibody Sampler Kit: Image 27
Western blot analysis of extracts from various cell lines using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 28
Immunoprecipitation of p300 from 293T cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p300 (D8Z4E) Rabbit mAb. Western blot analysis was performed using p300 (D8Z4E) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 29
CUT&RUN was performed with 293T cells and p300 (D8Z4E) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 1 (upper), including MCL1 (lower), a known target gene of p300 (see additional figure containing qPCR data).
Hypoxia Activation IHC Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GSK-3β (D5C5Z) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 31
Confocal immunofluorescent analysis of MCF-7 cells using LDHA (C4B5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Hypoxia Activation IHC Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human lymphoma using PKM2 (D78A4) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 33
Western blot analysis of extracts from NCI-H3122 cells, untreated (-) or treated with DMOG (1 mM, 6 hr; +), using HIF-1α (E1V6A) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Hypoxia Activation IHC Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using HIF-1β/ARNT (D28F3) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded human clear cell renal cell carcinoma using VHL (E3X9K) Rabbit mAb performed on the Leica BOND Rx.
Hypoxia Activation IHC Antibody Sampler Kit: Image 38
Confocal immunofluorescent analysis of MEF WT (left, positive) and MEF SirT1 KO (right, negative) cells usingSirT1 (1F3) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Wild-type and knockout MEFs were a gift from Wenyi Wei at the Harvard Medical School.
Hypoxia Activation IHC Antibody Sampler Kit: Image 39
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using p300 (D8Z4E) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 40
CUT&RUN was performed with 293T and either p300 (D8Z4E) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human MCL1 promoter primers, human MYC promoter primers, and human RSPh9 promoter primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Hypoxia Activation IHC Antibody Sampler Kit: Image 41
Immunohistochemical analysis of paraffin-embedded mouse pancreas using GSK-3β (D5C5Z) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 42
Immunohistochemical analysis of paraffin-embedded human skeletal muscle using PKM2 (D78A4) XP® Rabbit mAb. Note the lack of staining in the skeletal muscle cells which do not express PKM2 while vessels within the tissue stain positively.
Hypoxia Activation IHC Antibody Sampler Kit: Image 43
Western blot analysis of extracts from NCI-H28 and MOLT-4 cells using HIF-1α (E1V6A) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Hypoxia Activation IHC Antibody Sampler Kit: Image 44
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 45
Immunohistochemical analysis of paraffin-embedded mouse colon using HIF-1β/ARNT (D28F3) XP® Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 46
Immunohistochemical analysis of paraffin-embedded Hep G2 cell pellet (left, high-expressing) or MOLT-4 cell pellet (right, low-expressing) using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 47
Immunohistochemical analysis of paraffin-embedded human small cell carcinoma of the salivary gland using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 48
Confocal immunofluorescent analysis of HeLa (left) and C2C12 (right) cells using SirT1 (1F3) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Hypoxia Activation IHC Antibody Sampler Kit: Image 49
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using p300 (D8Z4E) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 50
Immunohistochemical analysis of paraffin-embedded MEF cell pellets, wild type (left), GSK-3α (-/-) (middle) and GSK-3β (-/-) (right) using GSK-3β (D5C5Z) XP® Rabbit mAb. (MEF wild type, GSK-3β (-/-), and GSK-3α (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Hypoxia Activation IHC Antibody Sampler Kit: Image 51
Confocal immunofluorescent analysis of A204 cells using PKM2 (D78A4) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Hypoxia Activation IHC Antibody Sampler Kit: Image 52
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 53
Chromatin immunoprecipitations were performed with cross-linked chromatin from T47D cells treated with BNF (1μM) for 45 min and HIF-1β/ARNT (D28F3) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)#9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across CYP1A2, a known target gene of HIF-1β (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Hypoxia Activation IHC Antibody Sampler Kit: Image 54
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 55
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin lymphoma using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 56
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 57
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using p300 (D8Z4E) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 58
Confocal immunofluorescent analysis of wild-type mouse embryonic fibroblasts (MEFs) (left), GSK-3α (-/-) MEFs (center)and GSK-3β (-/-) MEFs (right) using GSK-3β (D5C5Z) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild type, GSK-3α (-/-), and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Hypoxia Activation IHC Antibody Sampler Kit: Image 59
Flow cytometric analysis of 293T cells, transfected with PKM2 siRNA (blue) or mock transfected (green), using PKM2 (D78A4) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.
Hypoxia Activation IHC Antibody Sampler Kit: Image 60
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 61
Chromatin immunoprecipitations were performed with cross-linked chromatin from T47D cells treated with BNF (1μM) for 45 min and either HIF-1β/ARNT (D28F3) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human NFE2L2 Intron 1 Primers #81126, human CYP1A1 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Hypoxia Activation IHC Antibody Sampler Kit: Image 62
Immunohistochemical analysis of paraffin-embedded human oral squamous cell carcinoma using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 63
Immunohistochemical analysis of paraffin-embedded human serous adenocarcinoma of the ovary using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 64
Immunohistochemical analysis of paraffin-embedded 293T cell pellet (left, positive) or HCT-15 cell pellet (right, negative) using p300 (D8Z4E) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 65
Flow cytometric analysis of GSK-3β (-/-) MEFs (blue, negative) and wild type mouse embryonic fibroblasts (MEFs) (green, positive) using GSK-3β (D5C5Z) XP® Rabbit mAb or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. (MEF wild type and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Hypoxia Activation IHC Antibody Sampler Kit: Image 66
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 67
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 68
Immunohistochemical analysis of paraffin-embedded human T-cell lymphoma using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 69
Confocal immunofluorescent analysis of 293T cells (left, positive) and HCT-15 cells (right, negative) using p300 (D8Z4E) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Hypoxia Activation IHC Antibody Sampler Kit: Image 70
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 71
Immunohistochemical analysis of paraffin-embedded normal human liver using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 72
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 73
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma (left) and gastric carcinoma (right) using HIF-1α (E1V6A) Rabbit mAb (top) or a HIF-1α Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human HIF-1α. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Hypoxia Activation IHC Antibody Sampler Kit: Image 74
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 75

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using VHL (E3X9K) Rabbit mAb.

Hypoxia Activation IHC Antibody Sampler Kit: Image 76
Immunohistochemical analysis of paraffin-embedded normal human esophagus using HIF-1α (E1V6A) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Hypoxia Activation IHC Antibody Sampler Kit: Image 77
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Glut1 (E4S6I) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). 
Hypoxia Activation IHC Antibody Sampler Kit: Image 78
Immunohistochemical analysis of paraffin-embedded normal human testis using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 79
Immunohistochemical analysis of paraffin-embedded NCI-H3122 cell pellets, untreated (left) or treated with DMOG (1 mM, 6 hr; right), using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 80
Immunohistochemical analysis of paraffin-embedded normal human prostate using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 81
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 82
Immunohistochemical analysis of paraffin-embedded NCI-H28 cell pellet (left, positive) or MOLT-4 cell pellet (right, negative) using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 83
Immunohistochemical analysis of paraffin-embedded normal human spleen using
 Glut1 (E4S6I) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 84

Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using VHL (E3X9K) Rabbit mAb.

Hypoxia Activation IHC Antibody Sampler Kit: Image 85
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 86
Immunohistochemical analysis of paraffin-embedded Raji cell pellets, untreated (left) or treated with cobalt chloride (CoCl2) (100 μM, 4 hr; right), using HIF-1α (E1V6A) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 87
Immunohistochemical analysis of paraffin-embedded normal human placenta (left), small intestine (middle), or lung (right) using VHL (E3X9K) Rabbit mAb (top) or a VHL Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on human VHL. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Hypoxia Activation IHC Antibody Sampler Kit: Image 88
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 89
Immunohistochemical analysis of paraffin-embedded normal human placenta using VHL (E3X9K) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb XP® IgG Isotype Control #3900 (right).
Hypoxia Activation IHC Antibody Sampler Kit: Image 90
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 91
Immunohistochemical analysis of paraffin-embedded HeLa cell pellet (left, positive) or 786-O cell pellet (right, negative) using VHL (E3X9K) Rabbit mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 92
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using SirT1 (1F3) Mouse mAb.
Hypoxia Activation IHC Antibody Sampler Kit: Image 93
Immunohistochemical analysis of paraffin-embedded normal human testis using SirT1 (1F3) Mouse mAb.
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To Purchase # 43065
Cat. # Size Qty. Price
43065T		 1 Kit  (6 x 20 microliters)
$ 744

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
HIF-1α (E1V6A) Rabbit mAb 48085 20 µl
  • WB
  • IHC
  • IF
 H M 120 Rabbit IgG
HIF-1β/ARNT (D28F3) XP® Rabbit mAb 5537 20 µl
  • WB
  • IP
  • IHC
  • ChIP
 H M R Mk 87 Rabbit IgG
VHL (E3X9K) Rabbit mAb 81292 20 µl
  • IHC
 H M Rabbit IgG
p300 (D8Z4E) Rabbit mAb 86377 20 µl
  • WB
  • IP
  • IHC
  • IF
  • C&R
 H Mk 300 Rabbit IgG
SirT1 (1F3) Mouse mAb 8469 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R Mk 120 Mouse IgG1
GSK-3β (D5C5Z) XP® Rabbit mAb 12456 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Mk 46 Rabbit IgG
PKM2 (D78A4) XP® Rabbit mAb 4053 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Mk 60 Rabbit IgG
LDHA (C4B5) Rabbit mAb 3582 20 µl
  • WB
  • IHC
  • IF
 H Mk 37 Rabbit IgG
Glut1 (E4S6I) Rabbit mAb 73015 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R Mk 45-60 Rabbit IgG

Product Description

The Hypoxia Activation IHC Antibody Sampler Kit provides an economical means of detecting select components involved in the regulation of HIF-1α, select components regulated by HIF-1α, and HIF-1β/ARNT protein in formalin-fixed, paraffin-embedded tissue samples.

Specificity / Sensitivity

Each antibody in the Hypoxia Activation IHC Antibody Sampler Kit detects endogenous levels of its target protein. HIF-1α (E1V6A) Rabbit mAb does not cross-react with HIF-2α protein. Non-specific staining of skeletal and cardiac muscle has been observed by immunohistochemistry using VHL (E3X9K) Rabbit mAb. p300 (D8Z4E) Rabbit mAb does not cross-react with CBP protein. GSK-3β (D5C5Z) XP® Rabbit mAb does not cross-react with GSK-3α protein. PKM2 (D78A4) XP® Rabbit mAb does not cross-react with PKM1 protein. Glut1 (E4S6I) Rabbit mAb does not cross-react with Glut2, Glut3, or Glut4 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala475 of human HIF-1α protein, Ile479 of human HIF-1β/ARNT protein, Ser406 of human PKM2 protein, near the carboxy terminus of human p300 protein and human Glut1 protein, and corresponding to the sequence of human LDHA protein.
Monoclonal antibodies are also produced by immunizing animals with recombinant proteins specific to human VHL protein, representing the central region of human SirT1 protein, and specific to the carboxy terminus of human GSK-3β protein.

Background

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism, and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel-Lindau protein (VHL) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7). HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8). In addition, a chromosomal translocation leading to a TEL-ARNT fusion protein is associated with acute myeloblastic leukemia (9). Studies also found that ARNT/HIF-1β expression levels decrease significantly in pancreatic islets from patients with type 2 diabetes, suggesting that HIF-1β plays an important role in pancreatic β-cell function (10). CBP (CREB-binding protein) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (11,12). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (12). The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as class III histone deacetylases. The first discovered and best characterized of these genes is Saccharomyces cerevisiae SIR2, which is involved in silencing of mating type loci, telomere maintenance, DNA damage response, and cell aging (13). SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of many cellular processes, including apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. Targets of SirT1 include acetylated p53 (14,15), p300 (16), Ku70 (17), forkhead (FoxO) transcription factors (17,18), PPARγ (19), and the PPARγ coactivator-1α (PGC-1α) protein (20). Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulates glycogen synthesis in response to insulin (21). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3K/Akt cell survival pathway whose activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (22,23). Pyruvate kinase is a glycolytic enzyme that catalyzes the conversion of phosphoenolpyruvate to pyruvate. In mammals, the M2 isoform (PKM2) is expressed during embryonic development (24). Lactate dehydrogenase (LDH) catalyzes the interconversion of pyruvate and NADH to lactate and NAD+. The major form of LDH found in muscle cells is the A (LDHA) isozyme (25). Glucose transporter 1 (Glut1, SLC2A1) is a widely expressed transport protein that transports a number of different aldose sugars into cells (26,27).
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