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PhosphoSitePlus® Resource

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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous 10 Rabbit IgG
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Immunohistochemistry (Paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using DF1 (D8G6H) Rabbit mAb (IHC-Specific).

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Immunohistochemistry (Paraffin)

mmunohistochemical analysis of paraffin-embedded human ovarian carcinoma using SDF1 (D8G6H) Rabbit mAb (IHC-Specific).

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Immunohistochemistry (Paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using DF1 (D8G6H) Rabbit mAb (IHC-Specific).

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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

Product Usage Information

Application Dilutions
Immunohistochemistry (Paraffin) 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.

Specificity / Sensitivity

SDF1 (D8G6H) Rabbit mAb (IHC-Specific) recognizes endogenous levels of total SDF1 protein by immunohistochemistry on paraffin-embedded tissue sections. This antibody recognizes both SDF1α and SDF1β.


Species Reactivity: Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human SDF1 protein.

The stromal cell derived factor 1 (SDF1, CXCL12) is a small, pro-inflammatory chemoattractant cytokine that regulates leukocyte trafficking through interactions with its cognate 7-transmembrane G protein-coupled receptors (1). The SDF1 receptor, CXCR4, also serves as a coreceptor for the entry of human immunodeficiency virus into target cells (2). SDF1 may regulate homing and maintenance of CXCR4-expressing stem or progenitor cells, including embryonic and many somatic stem cells (3,4). Many cancer cells express CXCR4, suggesting that SDF1 plays a role in cancer metastasis (5,6). Alternative splicing and differential processing during maturation produce a pair of SDF1 isoforms (SDF1α and SDF1β) that have different properties and biological activities (7). Additional isoforms of SDF1 have been reported (8,9).


1.  Maekawa, T. and Ishii, T. (2000) Intern Med 39, 90-100.

2.  Ray, N. and Doms, R.W. (2006) Curr Top Microbiol Immunol 303, 97-120.

3.  Sugiyama, T. et al. (2006) Immunity 25, 977-88.

4.  Ara, T. et al. (2003) Immunity 19, 257-67.

5.  Kucia, M. et al. (2004) J Mol Histol 35, 233-45.

6.  Ratajczak, M.Z. et al. (2006) Leukemia 20, 1915-24.

7.  De La Luz Sierra, M. et al. (2004) Blood 103, 2452-9.

8.  Gleichmann, M. et al. (2000) Eur J Neurosci 12, 1857-66.

9.  Yu, L. et al. (2006) Gene 374, 174-9.


Entrez-Gene Id 6387
Swiss-Prot Acc. P48061


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.
Tween is a registered trademark of ICI Americas, Inc.

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SDF1 (D8G6H) Rabbit mAb (IHC-Specific)