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31549
Cancer Associated Fibroblast Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Cancer Associated Fibroblast Marker Antibody Sampler Kit #31549

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Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 1
Simple Western™ analysis of extracts (1 mg/mL) from Human Colon tissue using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb #19245. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 2
Simple Western™ analysis of lysates (0.1 mg/mL) from 3T3 cells using PDGF Receptor β (28E1) Rabbit mAb #3169. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 3
Immunoprecipitation of PDGF Receptor α from NCI H1703 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PDGF Receptor α (D1E1E) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 4
Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using Vimentin (D21H3) XP ® Rabbit mAb #5741. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 5
Western blot analysis of extracts from A172 and HeLa cells using S100A4 (D9F9D) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 6
Western blotting analysis of extracts from various human, mouse, and rat tissues using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (upper) and GAPDH #5174 (lower). As expected, skeletal muscle samples are negative for α-smooth muscle actin.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 7
Western blot analysis of extracts from various cell lines, using PDGF Receptor β (28E1) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 8
Western blot analysis of extracts from NIH/3T3 and human skeletal muscle cells (SKMC), untreated or treated with PDGF-BB, using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 9
Western blot analysis of extracts from various cell lines using Vimentin (D21H3) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 10
Western blot analysis of extracts from various cell lines using FAP (E1V9V) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 11
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 12
Immunoprecipitation of S100A4 from A172 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or S100A4 (D9F9D) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using S100A4 (D9F9D) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 13
Immunoprecipitation of α-smooth muscle actin from mouse colon tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb. Western blot analysis was performed using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PDGF Receptor β (28E1) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded human glioblastoma using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 16
Western blot analysis of extracts from control HeLa cells (lane 1) or Vimentin knockout HeLa cells (lane 2) using Vimentin (D21H3) XP® Rabbit mAb #5741 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Vimentin knockout HeLa cells confirms specificity of the antibody for Vimentin.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 17
Immunoprecipitation of FAP protein from M14 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is FAP (E1V9V) Rabbit mAb. Western blot analysis was performed using FAP (E1V9V) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 18
Immunohistochemical analysis of paraffin-embedded human colon using S100A4 (D9F9D) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded human endometrioid carcinoma using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb performed on the Leica® Bond™ Rx.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human glioblastoma using PDGF Receptor β (28E1) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded human colon using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 22
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 23
Confocal immunofluorescent analysis of U-138 MG cells (left, positive) or SK-MEL-28 cells (right, negative) using FAP (E1V9V) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using S100A4 (D9F9D) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded human appendix using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded U-87MG cells, showing membrane localization, using PDGF Receptor β (28E1) Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded U-118 MG xenograft using PDGF Receptor α (D1E1E) XP® Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 28
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Vimentin (D21H3) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 29
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded HCC827 xenograft using PDGF Receptor α (D1E1E) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Vimentin (D21H3) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 33
Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or PDGF-treated (right), using PDGF Receptor beta (28E1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 34
Confocal immunofluorescent analysis of A-204 (left) and U-87 MG cells (right) using PDGF Receptor α (D1E1E) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded mouse colon using Vimentin (D21H3) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded human tonsil using Vimentin (D21H3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 38
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 39
Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and IMR-32 cells (green, positive) using PDGF Receptor α (D1E1E) 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 (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 40
Confocal immunofluorescent analysis of SNB19 cells using Vimentin (D21H3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 41
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 42
Flow cytometric analysis of MCF7 cells (blue) and HeLa cells (green) using Vimentin (D21H3) 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 (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 43
Immunohistochemical analysis of paraffin-embedded mouse small intestine using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
Cancer Associated Fibroblast Marker Antibody Sampler Kit: Image 44
Confocal immunofluorescent analysis of mouse small intestine (left) and skeletal muscle (right) using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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To Purchase # 31549
Cat. # Size Qty. Price
31549T		 1 Kit  (6 x 20 microliters)
$ 518

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
PDGF Receptor α (D1E1E) XP® Rabbit mAb 3174 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M 190 Rabbit IgG
α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb 19245 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R 42 Rabbit IgG
PDGF Receptor β (28E1) Rabbit mAb 3169 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R 190 Rabbit IgG
Vimentin (D21H3) XP® Rabbit mAb 5741 20 µl
  • WB
  • IHC
  • IF
  • F
 H M R Mk 57 Rabbit IgG
FAP (E1V9V) Rabbit mAb 66562 20 µl
  • WB
  • IP
  • IF
 H 90 Rabbit IgG
S100A4 (D9F9D) Rabbit mAb 13018 20 µl
  • WB
  • IP
  • IHC
 H M 12 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Rab Goat 

Product Description

The Cancer Associated Fibroblast Marker Antibody Sampler Kit provides an economical means of detecting proteins reported to be expressed in Cancer Associated Fibroblasts (CAFs). The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb recognizes endogenous levels of total α-smooth muscle protein. PDGF Receptor α (D1E1E) XP® Rabbit mAb detects endogenous levels of PDGFRα. This antibody may cross-react with PDGFRβ at overexpressed levels. Nuclear staining has been observed with this antibody in certain tissues. The specificity of this staining is unknown. PDGF Receptor β (28E1) Rabbit mAb detects endogenous levels of PDGF receptor β protein. The antibody may cross-react with PDGF receptor α when highly overexpressed. Vimentin (D21H3) XP® Rabbit mAb detects endogenous levels of total vimentin protein. FAP (E1V9V) Rabbit mAb recognizes endogenous levels of total FAP protein. S100A4 (D9F9D) Rabbit mAb recognizes endogenous levels of total S100A4 protein.

Source / Purification

α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human α-smooth muscle actin protein. PDGF Receptor α (D1E1E) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy-terminal sequence of human PDGFRα. PDGF Receptor β (28E1) Rabbit mAb is produced by immunizing animals with a GST fusion protein containing a carboxy-terminal fragment of human PDGF receptor β. Vimentin (D21H3) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg45 of human vimentin protein. FAP (E1V9V) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala497 of human FAP protein. S100A4 (D9F9D) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala54 of human S100A4 protein.

Background

The tumor microenvironment (TME) has been shown to play an important role in tumor initiation, development, and metastasis. Numerous factors contribute to the nature of the TME such as the presence of immune cells; T-cells, B-cells, and natural killer (NK) cells, and wider environmental factors, such as extracellular matrix (ECM) stiffness, hypoxia, and interstitial pressure. Amongst all these various factors, fibroblasts have been suggested to play a key role in tumor development. 

Fibroblasts have been studied extensively, however, much regarding their influence on the TME remains to be understood. During tumor development, a subpopulation of hyper-activated fibroblasts become prominent in the TME and secretion of cytokines and chemokines from these cells promotes pro-tumorigenic activity. These highly heterogeneous fibroblast populations are known collectively as CAFs (Cancer Associated Fibroblasts).

Due to high plasticity and variability within CAF populations it has been difficult for researchers to define a universal marker for these cells. In lieu of a single marker, a number of markers are currently used to investigate CAFs. PDGFRα and PDGFRβ are common markers used for fibroblast identification, although PDGFRα is more widely expressed over the larger fibroblast populations. α-Smooth Muscle Actin is widely used to identify CAFs, however, some reports suggest it is not expressed by all functionally active CAFs. FSP-1/S100A4 is expressed by cells of mesenchymal origins. Although commonly used as a CAF marker, it too is not expressed by all fibroblasts present in a tumor. Some reports even suggest it to be a marker for quiescent fibroblasts. Fibroblast Activation Protein, or FAP as it is more commonly known, has traditionally been associated with tissue repair, fibrosis, and extracellular matrix degradation. FAP has more recently been described as a useful marker of CAFs. Vimentin strongly characterizes cells of a mesenchymal phenotype. It is frequently used as one marker of CAFs, but it is important to note that it is also highly expressed in fibroblasts of all types, as well as numerous other cell types, such as macrophages and adipocytes, and by epithelial cells undergoing epithelial-to-mesenchymal transition (EMT) (Reviewed in 1,2).
  1. LeBleu, V.S. and Kalluri, R. (2018) Dis Model Mech 11, pii: dmm029447. doi: 10.1242/dmm.029447.
  2. Nurmik, M. et al. (2019) Int J Cancer, doi: 10.1002/ijc.32193.

Pathways

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Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
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