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Collagen type IV antibody - 462 004

Collagen type IV is a basement membrane protein
Guinea pig polyclonal antiserum
Cat. No.: 462 004
Amount: 100 µl
Price: $365.00
Cat. No. 462 004 100 µl antiserum, lyophilized. For reconstitution add 100 µl H2O, then aliquot and store at -20°C until use.
Antibodies should be stored at +4°C when still lyophilized. Do not freeze!
Applications
 
WB: 1 : 1000 (AP-staining) gallery  
IP: not tested yet
ICC: not tested yet
IHC: 1 : 250 up to 1 : 1000 (see remarks) gallery  
IHC-P: 1 : 500 up to 1 : 100 (see remarks) gallery  
IHC-Fr: 1 : 500 up to 1 : 1000 (see remarks) gallery  
IHC-G: 1 : 500 up to 1 : 1000 (see remarks) gallery  

Western blot (WB); separation of proteins by PAGE and subsequent transfer to a membrane. Detection of target molecules is carried out with antibodies. Some antibodies require special sample preparation steps. For details, please refer to the “Remarks” section.

Immunoprecipitation (IP); Immunoisolation or pulldown of a target molecule using an antibody. For details and product specific hints, please refer to the ”Remarks” section.

Immunocytochemistry (ICC) on 4% PFA fixed cells. Immunoreactivity is usually revealed by fluorescence. Some antibodies require special fixation methods. For details, please refer to the “Remarks” section.

Immunohistochemistry (IHC) on 4% PFA perfusion fixed tissue with 24h PFA post fixation. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate. Some antibodies require special fixation methods or antigen retrieval steps. For details, please refer to the ”Remarks” section.

Immunohistochemistry (IHC-P) of formalin fixed, paraffin embedded (FFPE) tissue (some antibodies require special antigen retrieval steps, please refer to the ”Remarks” section). Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

Immunohistochemistry on fresh frozen (IHC-Fr) cryo-tissue-sections. In contrast to standard PFA perfusion fixed tissues, fresh frozen cryo-tissue-sections can be variably postfixed with alcohols, acetone or PFA. Alcohol or acetone fixation is e.g. of advantage for antigens masked by PFA crosslinking. For recommended postfixation, please refer to the ”Remarks” section. Immunoreactivity is usually revealed by fluorescence or a chromogenic substrate.

Immunohistochemistry on glyoxal fixed (IHC-G) tissue. The tissue is perfused with saline and afterwards immersion fixed with a glyoxal solution. For details of the glyoxal solution, please refer to the remarks section. Immunoreactivity is usually revealed by fluorescence.

Immunogen human heterotrimeric Collagen type IV containing two α 1-like (UniProt Id: P02462) and one α 2-like (UniProt Id: P08572) chains.
Reactivity Reacts with: mouse (P02463), rat (F1MA59), human (P02462), human (P08572), mouse (P08122), rat (F1M6Q3).
Other species not tested yet.
Remarks

IHC: antigen retrieval (0.2 mg/ml pepsin in 0.2 M HCl for 20 min at 37 °C) is recommended.
IHC-P: strong antigen retrieval is necessary
-  e.g.: antigen retrieval with Tris- EDTA buffer pH 9.0 in a food steamer followed with pepsin antigen retrieval or an antigen retrieval in a pressure cooker
IHC-Fr: PFA fixation is recommended.
IHC-G: Fixation with 3% glyoxal, 1% acetic acid, 20% ethanol in ddH2O according to Richter et al. 2017, or 9% glyoxal, 8% acetic acid in ddH2O according to Konno et al. 2023 are recommended.

Data sheet 462_004.pdf
Cat. No.: 462 004
Amount: 100 µl
Price: $365.00
Background

Type IV collagen is a unique member of the large collagen superfamily which in vertebrates comprises 28 different types. Unlike most collagens, type IV collagen occurs only in the basement membranes (BMs) and comprises up to six genetically distinct α-chains designated α1(IV) to α6(IV). Out of many potential combinations, the chains interact and assemble with a remarkable specificity to form only three distinct heterotrimers of α1α1α2, α3α4α5, and α5α5α6 (1). It has been shown that ablation of COL4 α 1/2 results in abnormal BM structure and embryonic lethality at E10.5–E11.5, although BM formation during early development is unaffected (2).