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47744
IGFBP7 (E3Z8L) XP® Rabbit mAb
Primary Antibodies

IGFBP7 (E3Z8L) XP® Rabbit mAb #47744

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  1. IHC

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using IGFBP7 (E3Z8L) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human kidney using IGFBP7 (E3Z8L) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using IGFBP7 (E3Z8L) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using IGFBP7 (E3Z8L) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using IGFBP7 (E3Z8L) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded mouse pancreas using IGFBP7 (E3Z8L) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded SCaBER cell pellet (left, positive) or HT-29 cell pellet (right, negative) using IGFBP7 (E3Z8L) XP® Rabbit mAb.

To Purchase # 47744S
Product # Size Price
47744T
20 µl $ 126
47744S
100 µl $ 302

Supporting Data

REACTIVITY H M
SENSITIVITY Endogenous
MW (kDa) 32
Isotype Rabbit IgG

Product Usage Information

Application Dilutions
Immunohistochemistry (Paraffin) 1:50 - 1:200

Storage:

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Protocol

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Immunohistochemistry (Paraffin)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. Xylene.
  2. Ethanol, anhydrous denatured, histological grade (100% and 95%).
  3. Deionized water (dH2O).
  4. Hematoxylin (optional).
  5. Wash Buffer:
    1. 1X Tris Buffered Saline with Tween® 20 (TBST): To prepare 1L 1X TBST add 100 ml 10X Tris Buffered Saline with Tween® 20 (#9997) to 900 ml dH20, mix.
  6. SignalStain® Antibody Diluent (#8112).
  7. 1X Citrate Unmasking Solution: To prepare 250 mL of 1X citrate unmasking solution, dilute 25 ml of SignalStain® Citrate Unmasking Solution (10X) (#14746) with 225 mL of dH2O.
  8. 3% Hydrogen Peroxide: To prepare 100 ml, add 10 ml 30% H2O2 to 90 ml dH2O.
  9. Blocking Solution: TBST/5% Normal Goat Serum or 1X Animal-Free Blocking Solution.
    1. TBST/5% Normal Goat Serum: to 5 ml 1X TBST, add 250 µl Normal Goat Serum (#5425).
    2. 1X Animal-Free Blocking Solution: to 4 mL of dH2O add 1 ml of Animal-Free Blocking Solution (5X) (#15019).
  10. Detection System: SignalStain® Boost IHC Detection Reagents (HRP, Rabbit #8114).
  11. Substrate: SignalStain® DAB Substrate Kit (#8059).
  12. Hematoxylin: Hematoxylin (#14166).
  13. Mounting Medium: SignalStain® Mounting Medium (#14177).

B. Deparaffinization/Rehydration

NOTE: Do not allow slides to dry at any time during this procedure.

  1. Deparaffinize/hydrate sections:
    1. Incubate sections in three washes of xylene for 5 min each.
    2. Incubate sections in two washes of 100% ethanol for 10 min each.
    3. Incubate sections in two washes of 95% ethanol for 10 min each.
  2. Wash sections two times in dH2O for 5 min each.

C. Antigen Unmasking

For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

D. Staining

  1. Wash sections in dH2O three times for 5 min each.
  2. Incubate sections in 3% hydrogen peroxide for 10 min.
  3. Wash sections in dH2O two times for 5 min each.
  4. Wash sections in wash buffer for 5 min.
  5. Block each section with 100–400 µl of preferred blocking solution for 1 hr at room temperature.
  6. Remove blocking solution and add 100–400 µl primary antibody diluted in SignalStain® Antibody Diluent (#8112) to each section. Incubate overnight at 4°C.
  7. Equilibrate SignalStain® Boost Detection Reagent (HRP, Rabbit #8114) to room temperature.
  8. Remove antibody solution and wash sections with wash buffer three times for 5 min each.
  9. Cover section with 1–3 drops SignalStain® Boost Detection Reagent (HRP, Rabbit #8114) as needed. Incubate in a humidified chamber for 30 min at room temperature.
  10. Wash sections three times with wash buffer for 5 min each.
  11. Add 1 drop (30 µl) SignalStain® DAB Chromogen Concentrate to 1 ml SignalStain® DAB Diluent and mix well before use.
  12. Apply 100–400 µl SignalStain® DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.
  13. Immerse slides in dH2O.
  14. If desired, counterstain sections with hematoxylin (#14166).
  15. Wash sections in dH2O two times for 5 min each.
  16. Dehydrate sections:
    1. Incubate sections in 95% ethanol two times for 10 sec each.
    2. Repeat in 100% ethanol, incubating sections two times for 10 sec each.
    3. Repeat in xylene, incubating sections two times for 10 sec each.
  17. Mount sections with coverslips and mounting medium (#14177).

posted February 2010

revised March 2016

Protocol Id: 283

Specificity / Sensitivity

IGFBP7 (E3Z8L) XP® Rabbit mAb recognizes endogenous levels of total IGFBP7 protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu241 of human IGFBP7 protein.

Background

IGFBP7 (also known as Mac25, TAF, or IGFBP-rP1) belongs to the IGFBP superfamily, which plays an integral role in regulating insulin-like growth factor (IGF) actions in a wide variety of cell types. There are six known high-affinity IGF binding family members (IGFBP1-6), and ten low-affinity IGF binding members. These family members are structurally related, but encoded by distinct genes (1,2). IGFBP7 is a low-affinity IGF binding protein (1). The protein functions through its binding to secreted growth factors including IGF1, insulin, and activin (3,4). IGFBP7 levels have been related to cancer development and tissue injury. Loss of expression of IGFBP7 has been associated with poor survival in multiple cancer types (5,6) and with tumor chemotherapy resistance (7,8). IGFBP7 also has been identified as a cell cycle arrest biomarker for human acute kidney injury (AKI) and serves as a prognostic indicator for early stage AKI development (9-11).

  1. Hwa, V. et al. (1999) Endocr Rev 20, 761-87.
  2. Duan, C. (2002) J Endocrinol 175, 41-54.
  3. Yamanaka, Y. et al. (1997) J Biol Chem 272, 30729-34.
  4. Kato, M.V. (2000) Mol Med 6, 126-35.
  5. Akiel, M. et al. (2014) J Hepatocell Carcinoma 1, 9-19.
  6. Benatar, T. et al. (2012) Breast Cancer Res Treat 133, 563-73.
  7. Verhagen, H.J. et al. (2014) Cell Death Dis 5, e1300.
  8. Okamura, J. et al. (2012) Cancer Biol Ther 13, 148-55.
  9. Kashani, K. et al. (2013) Crit Care 17, R25.
  10. Kellum, J.A. and Chawla, L.S. (2016) Nephrol Dial Transplant 31, 16-22.
  11. Gocze, I. et al. (2015) PLoS One 10, e0120863.

Pathways & Proteins

Explore pathways + proteins related to this product.

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.